P
US8932607B2ActiveUtilityPatentIndex 91

Batches of recombinant adenovirus with altered terminal ends

Assignee: CRUCELL HOLLAND BVPriority: Mar 12, 2012Filed: Mar 11, 2013Granted: Jan 13, 2015
Est. expiryMar 12, 2032(~5.7 yrs left)· nominal 20-yr term from priority
Inventors:CUSTERS JEROME H H VVELLINGA JORT
C12N 2710/10343C12N 2760/16234C12N 7/00C12N 2710/10321C12N 2760/16121C12N 2750/14143C12N 2760/16151C12N 2710/10351
91
PatentIndex Score
33
Cited by
135
References
46
Claims

Abstract

Described is a composition comprising a plurality of recombinant adenovirus particles, being a recombinant human adenovirus of serotype 5, 26, 34, 35, 48, 49 or 50, or a recombinant simian adenovirus, characterized in that the genomes of essentially all adenovirus particles in the composition comprise as the 5′ terminal nucleotides the nucleotide sequence: CTATCTAT (nucleotides 1-8 of SEQ ID NO:7). Also described are methods to produce such compositions.

Claims

exact text as granted — not AI-modified
What is claimed is: 
     
       1. A composition comprising adenovirus particles, wherein the adenovirus is a recombinant human adenovirus of serotype 5, 11a, 26, 34, 35, 48, 49 or 50, or a recombinant simian adenovirus, wherein the genomes of essentially all adenovirus particles in the composition have as the 5′ terminal nucleotides the polynucleotide: CTATCTAT, and wherein the adenovirus comprises a transgene. 
     
     
       2. The composition of  claim 1 , wherein the adenovirus is a human adenovirus of serotype 5, 26, 35, 49, or 50. 
     
     
       3. The composition of  claim 2 , wherein the adenovirus is a human adenovirus of serotype 26 or 35. 
     
     
       4. The composition of  claim 1 , which is a pharmaceutical composition. 
     
     
       5. The composition of  claim 1 , wherein the adenovirus lacks at least a portion of the E1 region. 
     
     
       6. The composition of  claim 1 , comprising at least 1×10 7  recombinant adenovirus particles. 
     
     
       7. A method for preparing a batch of recombinant adenovirus particles that have essentially all identical polynucleotides in the 5′ termini of their genomes, the method comprising:
 a) performing a molecular cloning step to exchange naturally occurring 5′ termini of an adenovirus genome with altered 5′ termini comprising as terminal nucleotides the polynucleotide: CTATCTAT, 
 b) propagating in host cells the recombinant adenovirus having the altered 5′ termini, and 
 c) harvesting the recombinant adenovirus to obtain a batch of recombinant adenovirus particles that essentially all have as the 5′ terminal nucleotides of their genomes the polynucleotide: CTATCTAT, and wherein the adenovirus comprises a transgene. 
 
     
     
       8. A method for preparing a batch of recombinant adenovirus particles that have essentially all identical polynucleotides in the 5′ termini of their genomes, the method comprising:
 a) performing a plaque purification of an adenovirus, wherein the recombinant adenovirus is a recombinant human adenovirus of serotype 5, 11a, 26, 34, 35, 48, 49 or 50, or a recombinant simian adenovirus, to isolate an adenovirus or recombinant adenovirus from a single plaque, wherein the adenovirus or recombinant adenovirus has as the 5′ terminal nucleotides of its genome polynucleotide: CTATCTAT, 
 b) propagating in host cells a recombinant adenovirus obtained from the single plaque of step a), and 
 c) harvesting the recombinant adenovirus to obtain a batch of recombinant adenovirus particles that essentially all have as the 5′ terminal nucleotides of their genomes the polynucleotide: CTATCTAT, and wherein the adenovirus comprises a transgene. 
 
     
     
       9. The method according to  claim 7 , wherein the batch comprises at least 1×10 7  recombinant adenovirus particles. 
     
     
       10. The method according to  claim 7 , wherein the recombinant adenovirus is a recombinant human adenovirus of serotype 5, 26, 35, 49, or 50. 
     
     
       11. The method according to  claim 7 , wherein the recombinant adenovirus is a recombinant human adenovirus of serotype 26 or 35. 
     
     
       12. The method according to  claim 7 , wherein the recombinant adenovirus lacks at least a portion of the E1 region. 
     
     
       13. The method according to  claim 7 , further comprising purifying the recombinant adenovirus. 
     
     
       14. The method according to  claim 13 , further comprising formulating the recombinant adenovirus into a pharmaceutical composition. 
     
     
       15. The method according to  claim 7 , wherein step b) is performed in a bioreactor. 
     
     
       16. The composition of  claim 6 , comprising at least 1×10 8  recombinant adenovirus particles. 
     
     
       17. The composition of  claim 16 , comprising at least 1×10 9  recombinant adenovirus particles. 
     
     
       18. The composition of  claim 17 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       19. The composition of  claim 1 , wherein the adenovirus is a human adenovirus of serotype 35. 
     
     
       20. The composition of  claim 1 , wherein the adenovirus is a human adenovirus of serotype 26. 
     
     
       21. The composition of  claim 2 , which is a pharmaceutical composition. 
     
     
       22. The composition of  claim 3 , which is a pharmaceutical composition. 
     
     
       23. The composition of  claim 19 , which is a pharmaceutical composition. 
     
     
       24. The composition of  claim 20 , which is a pharmaceutical composition. 
     
     
       25. The composition of  claim 2 , wherein the adenovirus lacks at least a portion of the E1 region. 
     
     
       26. The composition of  claim 3 , wherein the adenovirus lacks at least a portion of the E1 region. 
     
     
       27. The composition of  claim 19 , wherein the adenovirus lacks at least a portion of the E1 region. 
     
     
       28. The composition of  claim 20 , wherein the adenovirus lacks at least a portion of the E1 region. 
     
     
       29. The composition of  claim 23 , wherein the adenovirus lacks at least a portion of the E1 region. 
     
     
       30. The composition of  claim 24 , wherein the adenovirus lacks at least a portion of the E1 region. 
     
     
       31. The composition of  claim 2 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       32. The composition of  claim 3 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       33. The composition of  claim 4 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       34. The composition of  claim 5 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       35. The composition of  claim 19 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       36. The composition of  claim 20 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       37. The composition of  claim 21 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       38. The composition of  claim 22 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       39. The composition of  claim 23 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       40. The composition of  claim 24 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       41. The composition of  claim 25 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       42. The composition of  claim 26 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       43. The composition of  claim 27 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       44. The composition of  claim 28 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       45. The composition of  claim 29 , comprising at least 1×10 10  recombinant adenovirus particles. 
     
     
       46. The composition of  claim 30 , comprising at least 1×10 10  recombinant adenovirus particles.

Cited by (0)

No later patents cite this yet.

References (0)

No backward citations on record.